Osawa Hiroki, Matsumoto Hideaki
Research Institute for Bioresources, Okayama University, 2-20-1 Chuo, Kurashiki, 710-0046 Japan.
Planta. 2006 Jul;224(2):462-71. doi: 10.1007/s00425-006-0220-5. Epub 2006 Feb 1.
Aluminum (Al) tolerance in wheat (Triticum aestivum L.) is mainly achieved by malate efflux, which is regulated by the expression of the recently identified gene, presumably encoding an Al-activated malate efflux transporter (ALMT1). However, the transport mechanism is not fully understood, partly as a result of the rapid turnover of its substrate. We developed a tool to study malate transport in wheat by screening biological compounds using the well-characterized Schizosaccharomyces pombe malate transporter (SpMAE1). Expression of SpMAE1 in both S. pombe and Saccharomyces cerevisiae, which has no SpMAE1 homologue, caused hypersensitivity to thio-malic acid. This hypersensitivity was prominent at pH 3.5, but not pH 4.5, and was accompanied by an increase in thiol content, indicating that SpMAE1 mediates the uptake of thio-malic acid at a specific low pH. In wheat, root apices were able to accumulate thio-malic acid without growth reduction at pH values above 4.2. Pretreatment of root apices with thio-malic acid followed by Al treatment induced thio-malate efflux. Al-induced thio-malate efflux was much higher in Al-resistant cultivars/genotypes than in Al-sensitive ones, and was accompanied by a decrease in thiol-content. Thio-malate efflux in the Al-resistant cultivar was slightly activated by lanthanum or ytterbium ion. Thio-malic acid did not alleviate the Al-induced inhibition of root elongation in wheat. Taken together, our results suggest that thio-malate acts as an analogue for malate in malate transport systems in wheat and yeast, and that it may be a useful tool for the analysis of malate transport involved in Al-tolerance and of other organic ion transport processes.
小麦(Triticum aestivum L.)对铝(Al)的耐受性主要通过苹果酸外流来实现,这一过程受最近鉴定出的基因表达调控,该基因可能编码一种铝激活的苹果酸外流转运蛋白(ALMT1)。然而,其转运机制尚未完全明确,部分原因是其底物周转迅速。我们开发了一种工具,通过使用特征明确的粟酒裂殖酵母苹果酸转运蛋白(SpMAE1)筛选生物化合物来研究小麦中的苹果酸转运。SpMAE1在粟酒裂殖酵母和没有SpMAE1同源物的酿酒酵母中表达,都会导致对硫代苹果酸过敏。这种过敏在pH 3.5时很明显,但在pH 4.5时不明显,并且伴随着硫醇含量的增加,这表明SpMAE1在特定的低pH值下介导硫代苹果酸的摄取。在小麦中,根尖在pH值高于4.2时能够积累硫代苹果酸而不影响生长。用硫代苹果酸预处理根尖后再进行铝处理会诱导硫代苹果酸外流。铝诱导的硫代苹果酸外流在耐铝品种/基因型中比在铝敏感品种中要高得多,并且伴随着硫醇含量的降低。耐铝品种中的硫代苹果酸外流会被镧或镱离子轻微激活。硫代苹果酸并不能缓解铝诱导的小麦根伸长抑制。综上所述,我们的结果表明硫代苹果酸在小麦和酵母的苹果酸转运系统中作为苹果酸的类似物起作用,并且它可能是分析参与铝耐受性的苹果酸转运和其他有机离子转运过程的有用工具。